Regulation of the nuclear factor of activated T cells in stably transfected Jurkat cell clones.
Regulation of the nuclear factor of activated T cells in stably transfected Jurkat cell clones.
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稳定转染的 Jurkat 细胞克隆中活化 T 细胞核因子的调节。
DOI：
10.1006/bbrc.1996.0187
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发表时间：
1996
影响因子：
3.1
通讯作者：
Handschumacher,RE
中科院分区：
文献类型：
--
作者：
Li,W;Handschumacher,RE
两个Jurkat细胞克隆已稳定转染的报告载体的核因子活化的T细胞（NFAT）。在刺激下，它们表达高水平的分泌型热稳定胎盘碱性磷酸酶。通过这些克隆，我们证明了由佛波醇12-肉豆蔻酸酯13-乙酸酯和离子霉素诱导的NFAT激活被环孢菌素A（CsA）（IC 50 = 8 nM）和FK 506（IC 50 = 160 pM）抑制，推测是通过抑制钙调磷酸酶活性。不抑制钙调磷酸酶的蛋白磷酸酶1（PP 1）和2A（PP 2A）的选择性磷酸酶抑制剂，如冈田酸和calyculin A，也分别以87 nM和4 nM的IC 50抑制NFAT活化，表明冈田酸和相关抑制剂可能通过抑制PP 1而不是PP 2A来阻断NFAT活化。NFAT活化也被增加cAMP浓度的试剂如二丁酰cAMP、毛喉素和前列腺素E2抑制。这些稳定的Jurkat细胞克隆被证明是研究NFAT调控的方便和灵敏的工具。
Two Jurkat cell clones have been stably transfected with a reporter vector for the nuclear factor of activated T cells (NFAT). Upon stimulation, they express high levels of secreted heat stable placental alkaline phosphatase. With these clones, we demonstrated that NFAT activation induced by phorbol 12-myristate 13-acetate and ionomycin was inhibited by both cyclosporin A (CsA) (IC50= 8 nM) and FK506 (IC50= 160 pM), presumably by inhibition of calcineurin activity. Selective phosphatase inhibitors for protein phophatase 1 (PP1) and 2A (PP2A) that do not inhibit calcineurin, such as okadaic acid and calyculin A, also inhibited NFAT activation with IC50s of 87 nM and 4 nM, respectively, suggesting that okadaic acid and related inhibitiors may block NFAT activation through the inhibition of PP1, instead of PP2A. NFAT activation was also inhibited by agents that increase cAMP concentrations such as dibutyryl cAMP, forskolin and prostaglandin E2. These stable Jurkat cell clones proved a convenient and sensitive tool to study NFAT regulation.